LOCUS       sequence-254617-        3198 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BTK Type 8 Bacterial origin (R6K suicide plasmid) for homologous 
            recombination.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3198)
  AUTHORS   Leonard SP, Perutka J, Powell JE, Geng P, Richhart DD, Byrom M, Kar 
            S, Davies BW, Ellington AD, Moran NA, Barrick JE
  TITLE     Genetic Engineering of Bee Gut Microbiome Bacteria with a Toolkit 
            for Modular Assembly of Broad-Host-Range Plasmids.
  JOURNAL   ACS Synth Biol. 2018 May 18;7(5):1279-1290. doi: 
            10.1021/acssynbio.7b00399. Epub 2018 Apr 13.
  PUBMED    29608282
REFERENCE   2  (bases 1 to 3198)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.7b00399"; journalName: "ACS Synth Biol"; date: 
            "2018-05-18- 18"; volume: "7"; issue: "5"; pages: "1279-1290"
COMMENT     Sequence Label: pBTK599
FEATURES             Location/Qualifiers
     source          1..3198
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             83..94
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             101..814
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      826..897
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      913..940
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     oriT            1056..1165
                     /note="incP origin of transfer"
     rep_origin      1336..1724
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     primer_bind     1757..1773
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1757..1773
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1807..1823)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1808..1824)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             complement(join(2127..2918,2919..2987))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2750..2769
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2988..3092)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gcaattcaca tttaatttat gaatgttttc ttaacatcgc ggcaactcaa gaaacggcag
       61 gttcggatct tagctactag agaaagagga gaaatactag atgcgtaaag gcgaagagct
      121 gttcactggt gtcgtcccta ttctggtgga actggatggt gatgtcaacg gtcataagtt
      181 ttccgtgcgt ggcgagggtg aaggtgacgc aactaatggt aaactgacgc tgaagttcat
      241 ctgtactact ggtaaactgc cggttccttg gccgactctg gtaacgacgc tgacttatgg
      301 tgttcagtgc tttgctcgtt atccggacca tatgaagcag catgacttct tcaagtccgc
      361 catgccggaa ggctatgtgc aggaacgcac gatttccttt aaggatgacg gcacgtacaa
      421 aacgcgtgcg gaagtgaaat ttgaaggcga taccctggta aaccgcattg agctgaaagg
      481 cattgacttt aaagaggacg gcaatatcct gggccataag ctggaataca attttaacag
      541 ccacaatgtt tacatcaccg ccgataaaca aaaaaatggc attaaagcga attttaaaat
      601 tcgccacaac gtggaggatg gcagcgtgca gctggctgat cactaccagc aaaacactcc
      661 aatcggtgat ggtcctgttc tgctgccaga caatcactat ctgagcacgc aaagcgttct
      721 gtctaaagat ccgaacgaga aacgcgatca tatggttctg ctggagttcg taaccgcagc
      781 gggcatcacg catggtatgg atgaactgta caaatgacca ggcatcaaat aaaacgaaag
      841 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac
      901 tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttataagtc cgtctcaagc
      961 aagggggatc aattccgtga taggtgggct gcccttcctg gttggcttgg tttcatcagc
     1021 catccgcttg ccctcatctg ttacgccggc ggtagccggc cagcctcgca gagcaggatt
     1081 cccgttgagc accgccaggt gcgaataagg gacagtgaag aaggaacacc cgctcgcggg
     1141 tgggcctact tcacctatcc tgcccggctg acgccgttgg atacaccaag gaaagtctac
     1201 acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag gatggatata ccgaaaaaat
     1261 cgctataatg accccgaagc agggttatgc agcggaaaac ggaattgatc cggccacgat
     1321 gcgtccggcg tagaggatct gaagatcagc agttcaacct gttgatagta cgtactaagc
     1381 tctcatgttt cacgtactaa gctctcatgt ttaacgtact aagctctcat gtttaacgaa
     1441 ctaaaccctc atggctaacg tactaagctc tcatggctaa cgtactaagc tctcatgttt
     1501 cacgtactaa gctctcatgt ttgaacaata aaattaatat aaatcagcaa cttaaatagc
     1561 ctctaaggtt ttaagtttta taagaaaaaa aagaatatat aaggctttta aagcttttaa
     1621 ggtttaacgg ttgtggacaa caagccaggg atgtaacgca ctgagaagcc cttagagcct
     1681 ctcaaagcaa ttttgagtga cacaggaaca cttaacggct gacatgggaa ttcccctcca
     1741 ccgcggtggc ggccgctcta gaactagtgg atcccccggg ctgcaggaat tcgatatcaa
     1801 gcttatcgat accgtcgacc tcgagggggg gcccggtacc gaggacgcgt cgaataattc
     1861 cgctagcttc acgctgccgc aagcactcag ggcgcaaggg ctgctaaagg aagcggaaca
     1921 cgtagaaagc cagtccgcag aaacggtgct gaccccggat gaatgtcagc tactgggcta
     1981 tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt agcttgcagt gggattatca
     2041 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2101 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2161 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     2221 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcggga cccacgctca
     2281 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     2341 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     2401 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     2461 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     2521 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     2581 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     2641 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     2701 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     2761 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     2821 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     2881 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     2941 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3001 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3061 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgcc
     3121 tgatgagacg atgaaagtga aacgtgattt catgcgtcat tttgaacatt ttgtaaatct
     3181 tatttaataa tgtgtgcg
//
